本文選題：人臍帶血 + 樹(shù)突狀細(xì)胞��； 參考：《第三軍醫(yī)大學(xué)》2006年碩士論文

【摘要】： 樹(shù)突狀細(xì)胞(dendritic cell, DC)是一類重要的專職抗原呈遞細(xì)胞(antigen-presenting cell , APC),雖然在體內(nèi)的數(shù)量較少,但是其強(qiáng)大的抗原遞呈和處理功能在機(jī)體的免疫反應(yīng)中起重要的作用。DC的發(fā)育分化過(guò)程伴隨著DC由不成熟的前體細(xì)胞向成熟細(xì)胞轉(zhuǎn)變,未成熟DC(immature dendritic cell, imDC)與成熟DC(mature dendritic cell, mDC)在表型特征以及生物學(xué)功能上都有區(qū)別,而imDC最大的特點(diǎn)就是在體外可以誘導(dǎo)T淋巴細(xì)胞特異性低應(yīng)答。臨床上同種異體皮膚移植是目前大面積深度燒傷患者早期創(chuàng)面覆蓋最直接、有效的治療方法,但是由于皮膚的強(qiáng)烈抗原特性,導(dǎo)致移植后平均3周左右外源皮膚就會(huì)發(fā)生不可逆的排斥反應(yīng),極大抑制了自體微粒皮混合大張異體皮移植效果。免疫抑制藥物雖可減輕免疫排斥,但是由于大面積嚴(yán)重?zé)齻颊叩拿庖吖δ芎慕?可導(dǎo)致嚴(yán)重感染威脅患者生命。若能有效利用imDC的特殊作用,移植前后在受者體內(nèi)輸入基因工程制備的imDC,可特異性地減輕皮膚移植后受者對(duì)供者抗原的免疫排斥反應(yīng)、延長(zhǎng)異體皮的存活時(shí)間,從而提高大面積深度燒傷患者的手術(shù)治療效果。 隨著基因治療的不斷深入和發(fā)展,利用各種基因工程改造DC誘導(dǎo)器官移植耐受的實(shí)驗(yàn)性研究已經(jīng)逐漸深入,如將各種趨化因子受體(chemokine receptor, CCR)的基因?qū)雐mDC中,使其靶向歸巢至引流淋巴結(jié)中,有效發(fā)揮誘導(dǎo)耐受的功能,或者將針對(duì)細(xì)胞毒性T淋巴細(xì)胞相關(guān)抗原免疫球蛋白(CTLA-4Ig)的基因?qū)雐mDC中,表達(dá)的產(chǎn)物抑制imDC和T細(xì)胞表面CTLA-4的結(jié)合從而抑制共刺激效應(yīng)和免疫激活,等等。因此,有效地實(shí)現(xiàn)imDC的免疫耐受功能需要將目的基因通過(guò)特定的方式整合到樹(shù)突狀細(xì)胞中,但在轉(zhuǎn)染過(guò)程中,存在的一個(gè)問(wèn)題是不同的轉(zhuǎn)染方式常能影響DC作為抗原遞呈細(xì)胞的功能,轉(zhuǎn)染未成熟樹(shù)突狀細(xì)胞后可能會(huì)誘導(dǎo)其成熟。 在本研究中,我們聯(lián)合應(yīng)用rhGM-CSF和rhIL-4體外誘導(dǎo)、擴(kuò)增人臍帶血分離的單核細(xì)胞,第7d收獲細(xì)胞,從形態(tài)學(xué)、細(xì)胞表面標(biāo)志物以及混合淋巴細(xì)胞反應(yīng)(MLR)中對(duì)同種異體未致敏T淋巴細(xì)胞刺激能力三個(gè)方面進(jìn)行鑒定。然后我們用兩種常用的
[Abstract]:Dendritic cell (dendritic cell, DC) is an important class of specialized antigen presenting cells (antigen-presenting cell, APC), although the number in vivo is relatively small. However, its powerful antigen presentation and processing function plays an important role in the immune response of the body. The development and differentiation of DC is accompanied by the transformation of DC from immature precursor cells to mature cells. The phenotypic characteristics and biological functions of immature DC (immature dendritic cell, MDC and mature DC (mature dendritic cell, mDC) are different, and the biggest characteristic of imDC is that it can induce T lymphocyte specific low response in vitro. Skin allograft transplantation is currently the most direct and effective treatment for the early wound coverage in patients with extensive deep burns, but due to the strong antigenic characteristics of the skin, This resulted in irreversible rejection of the skin after transplantation for an average of 3 weeks, which greatly inhibited the effect of autologous skin grafts mixed with large skin allografts. Immunosuppressive drugs can reduce the immune rejection, but due to the exhaustion of immune function in patients with severe burn, it can lead to serious infection and threaten the lives of patients. If the special function of imDC can be effectively utilized, imDCs prepared by gene engineering before and after transplantation can specifically alleviate the immune rejection of donor antigen and prolong the survival time of allogeneic skin after skin transplantation. So as to improve the surgical treatment of large area deep burn patients. With the development of gene therapy, the experimental research on the induction of organ transplantation tolerance by using various genetic engineering has been gradually deepened, such as the introduction of various chemokine receptor (chemokine receptor,) genes into imDC. By targeting homing to the draining lymph nodes, it can effectively induce tolerance, or transfer the CTLA-4Ig gene into IMDC, which is related to the cytotoxic T lymphocyte antigen (CTLA-4Ig). The expressed product inhibits the binding of imDC to CTLA-4 on T cell surface, which inhibits costimulatory effect and immune activation, and so on. Therefore, it is necessary to integrate the target gene into dendritic cells in a specific way to achieve the immune tolerance function of imDC, but in the process of transfection, One problem is that different transfection methods can often affect the function of DC as antigen presenting cells, and immature dendritic cells may be induced to mature after transfection. In this study, we combined rhGM-CSF and rhIL-4 to amplify monocytes isolated from human umbilical cord blood in vitro. Cell surface markers and the stimulating ability of allogeneic unsensitized T lymphocytes were identified in mixed lymphocyte reaction (MLR). And then we use two commonly used
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 王強(qiáng),彭毅志;小鼠骨髓未成熟樹(shù)突狀細(xì)胞體外擴(kuò)增及鑒定[J];中華燒傷雜志;2003年06期

2 鄭峻松,吳軍,肖光夏;樹(shù)突狀細(xì)胞與移植免疫耐受的研究[J];中華燒傷雜志;2003年06期

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本文編號(hào)：2054822